BHLHB2 在人胰腺癌细胞株SU86.86的生物学功能研究

 【摘要】目的：进一步研究前期实验结果证明的人胰腺癌新颖的独立预后因子BHLHB2(cAMP -inducible basic helix-loop-helix domain containing, class B, 2) 在人胰腺癌细胞中的相关分子生物学功能。方法：应用siRNA干扰技术，在人胰腺癌细胞株SU86.86中使BHLHB2表达沉寂，然后进行迁徙实验，研究BHLHB2与细胞迁徙的关系；应用更生霉素（Actinomycin D，ACT-D)诱导细胞凋亡，研究BHLHB2与细胞凋亡的关系；应用Western Blot方法研究BHLHB2与EMT的相关性。结果: 人胰腺癌细胞BHLHB2 表达沉寂后，细胞的迁徙明显增快。应用ACT-D进行凋亡诱导，BHLHB2表达沉寂后、发生凋亡的细胞数目明显增加(10ng/ml, 增加 1.32倍, p<0.05; 100ng/ml, 增加 1.41倍, p<0.05)。Western Blot结果表明, BHLHB2表达沉寂后可显著降低E-cadherin的表达(降低 11.35 倍, p<0.01)。结论: BHLHB2 具有多种生物学功能，与人胰腺癌细胞的迁徙、细胞凋亡及EMT密切相关。

BHLHB2 participates in cell migration,cell apoptosis and EMT in human pancreatic cancer

Objective To further explore BHLHB2 related molecular biology functions,which was proved to be a novel and independent prognostic marker in pancreatic ductal adenocarcinoma.Methods Knock-down of BHLHB2 in pancreatic cancer cell line-SU86.86 was achieved by siRNA treatment.Migration essay was performed to research the relationship between BHLHB2 and cell migration.The proliferation assay induced by ACT-D to explore BHLHB2 roles in cell apoptosis.Relationship between BHLHB2 and EMT was analyzed using Western blot.Results BHLHB2 silencing with RNAi significantly promote cell migrate,and induced cell apoptosis by Actinomycin D(ACT-D)(10 μg/L,increased 1.32 fold,P<0.05;100 μg/L,increased 1.41 fold,P<0.05).By Western blot,BHLHB2 silencing significantly deduced E-cadherin expression(decreased 1.28 fold,P<0.01),which implicated that BHLHB2 was correlated with epithelial mesenchymal transform(EMT).Conclusions BHLHB2 closely associated with cell migration,cell apoptosis and EMT in human pancreatic cancer.